Profiling of Histone Post-Translational Modifications in Mouse Brain with High-Resolution Top-Down Mass Spectrometry
Mowei Zhou1, Ljiljana Paša-Tolić1, David L Stenoien1
1Pacific Northwest National Laboratory, Earth and Biological Sciences Directorate , P.O. Box 999, Richland, Washington 99352, United States.
Journal of Proteome Research
|December 22, 2016
Summary
Histone post-translational modifications (PTMs) were analyzed using advanced mass spectrometry. This study identified novel histone PTMs, including tyrosine bromination and glutathionylation, advancing proteoform characterization.
Area of Science:
- Biochemistry
- Molecular Biology
- Proteomics
Background:
- Histones are crucial for DNA replication, repair, and transcription.
- Histone post-translational modifications (PTMs) regulate these functions.
- Complete characterization of histone proteoforms requires top-down analysis, but faces technical challenges.
Purpose of the Study:
- To develop and apply a robust workflow for comprehensive top-down histone analysis.
- To identify novel histone PTMs and map known modifications across entire histone proteins.
Main Methods:
- Utilized state-of-the-art mass spectrometry.
- Implemented a bioinformatics workflow including ProMex, MSPathFinder, LcMsSpectator, and TopPIC.
- Employed targeted data analysis and visualization for intact protein analysis.
Main Results:
- Successfully identified novel histone PTMs, such as tyrosine bromination on histones H4 and H2A.
- Discovered glutathionylation on histone H3.
- Mapped conventional PTMs across entire histone subunits, enhancing proteoform understanding.
Conclusions:
- The developed workflow significantly improves top-down histone analysis capabilities.
- This research expands the known landscape of histone PTMs, offering new insights into their regulatory roles.
- Advanced proteomic strategies are essential for dissecting complex histone modifications and their impact on cellular functions.
